CN108479110A - A kind of cell culture liquid decolorizer and discoloration method - Google Patents
A kind of cell culture liquid decolorizer and discoloration method Download PDFInfo
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- CN108479110A CN108479110A CN201810315766.7A CN201810315766A CN108479110A CN 108479110 A CN108479110 A CN 108479110A CN 201810315766 A CN201810315766 A CN 201810315766A CN 108479110 A CN108479110 A CN 108479110A
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/14—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the introduction of the feed to the apparatus
Abstract
The present invention provides a kind of cell culture liquid decolorizer and discoloration methods, wherein the cell culture liquid decolorizer includes：It is one or more in chitosan, polyaluminium chloride and newborn calcium phosphate；Wherein, the concentration control of each component is in cell culture fluid：100 2000mg/L of chitosan；50 150mg/L of polyaluminium chloride；20 150mM of newborn calcium phosphate.The cell culture liquid decolorizer is realized in the production of the biogenetic products such as monoclonal antibody or diagnostic reagent, abundant purifying and decoloration to pigment and impurity in cell culture fluid, purification efficiency is high, burden is alleviated the work such as further to extract, filtering, the cost for greatly reducing purifying and production brings huge convenience to the production, research and development and use of cell culture fluid.
Description
Technical field
The invention belongs to biotechnology more particularly to a kind of cell culture liquid decolorizers and discoloration method.
Background technology
With the fast development of biotechnology, mammalian cell is widely used in including monoclonal antibody medicine, diagnosis examination
The production of the biological products such as agent plays an important role to human medical's health.
Monoclonal antibody medicine, diagnostic reagent are needed in cell culture fluid by clarification extraction.However it is trained in cell
In order to improve the quality of yield and antibody during supporting, it usually needs add various nutriments, cause in culture solution
A large amount of accumulation pigments, increase the burden of purification step, particularly with diagnostic reagent, there are the spirits that a large amount of pigment can interfere diagnosis
Sensitivity.
Currently, in monoclonal antibody production especially diagnostic reagent production, first centrifugation is mostly used greatly or by filter with high costs
Material filters, and then is extracted again by protein A or other kinds of prepacked columns, since there are conventional purifications in culture solution
Much in the impurity and pigment that can not be purified in technology, cause for further protein A or other kinds of prepacked columns
Service life cause larger damage and can not regeneration, purifying is with high costs, influences the quality of finished product, to cell culture fluid
Production, research and development and use bring huge inconvenience.
Invention content
A kind of cell culture liquid decolorizer of present invention offer and discoloration method, to solve the deficiency in existing technology.
To solve the above problems, the present invention provides a kind of cell culture liquid decolorizer, including：
It is one or more in chitosan, polyaluminium chloride and newborn calcium phosphate；
Wherein, the concentration control of each component is in cell culture fluid：
Chitosan 100-2000mg/L；
Polyaluminium chloride 50-150mg/L；
Newborn calcium phosphate 20-150mM.
Preferably, the concentration control of each component is in the cell culture fluid：
Chitosan 500-1500mg/L；
Polyaluminium chloride 80-100mg/L；
Newborn calcium phosphate 40-120mM.
Preferably, further include cosolvent；
The cosolvent is concentrated hydrochloric acid；
Final concentration control of the cosolvent in the cell culture fluid is 0.001-0.1mg/L.
In addition, to solve the above problems, the present invention also provides a kind of cell culture fluid discoloration methods, including：
Chitosan is subjected to hydrotropy dissolving by cosolvent, obtains chitosan solution；
Newborn calcium phosphate is prepared when carrying out culture solution decoloration；Then, the chitosan solution, polyaluminium chloride and new life are taken
The one or more of calcium phosphate are separately added into the culture solution to be decolourized；Wherein, it is poly- that the newborn calcium phosphate, shell are prepared respectively
Sugared, polyaluminium chloride amount obtains mixed-culture medium to reach default final concentration；
The mixed-culture medium is mixed and is centrifuged to get to the cell culture fluid after decoloration.
Preferably, described " chitosan being carried out hydrotropy dissolving by cosolvent, obtain chitosan solution " includes：
Based on default chitosan solution concentration, 0.1-5 parts of the chitosan is added to the water mixing, obtains concentrating molten
Liquid；
0.001-0.1 parts of the cosolvent is added in the concentrate solution to get to chitosan solution.
Preferably, a concentration of 3-10g/L of default chitosan solution.
Preferably, described " newborn calcium phosphate is prepared when carrying out culture solution decoloration ", including：
It takes calcium chloride and sodium dihydrogen phosphate to be added to the water reaction and generates calcium phosphate, the newborn phosphorus is prepared after mixing
Sour calcium.
Preferably, described default final concentration of：
Chitosan 100-2000mg/L；
Polyaluminium chloride 50-150g/L；
Calcium phosphate 40-120mM.
Preferably, described " mixed-culture medium is mixed and is centrifuged to get to the cell culture fluid after decoloration "
Including：
The mixed solution is stirred 5-10 minutes at ambient temperature, mixing speed 50-100rpm is stirred
Mix mixed solution；
To it is described be stirred solution carry out centrifugation centrifugal force be set as 8000-12000g, 2-8 DEG C of temperature, time 20-30
Minute to get to the cell culture fluid after the decoloration.
A kind of cell culture liquid decolorizer of present invention offer and discoloration method.Wherein, the cell culture liquid decolorizer is logical
It crosses using the obtained cell culture liquid decolorizer of one or more combinations in chitosan, polyaluminium chloride and newborn calcium phosphate,
It is acted on using the flocculation adsorption of chitosan, polyaluminium chloride and newborn calcium phosphate, by adding chitosan in cell culture fluid, gathering
One or more combinations in aluminium chloride and newborn calcium phosphate carry out adsorpting pigment, and then by pigment as cell removes together, from
And reach decolorizing effect, to realize in the production of the biogenetic products such as monoclonal antibody or diagnostic reagent, to the color in cell culture fluid
The abundant purifying and decoloration of element and impurity, purification efficiency is high, alleviates burden for work such as further extraction, filterings, significantly
The cost for reducing purifying and production brings huge convenience to the production, research and development and use of cell culture fluid.
Specific implementation mode
Technical scheme of the present invention is described in further detail with reference to the mode of specific embodiment, but not structure
At any limitation of the invention, the modification of anyone limited number of time made within the scope of the invention as claimed, still in this hair
Within bright right.
The present invention provides a kind of cell culture liquid decolorizer, including：
It is one or more in chitosan, polyaluminium chloride and newborn calcium phosphate；
Wherein, the concentration control of each component is in cell culture fluid：
Chitosan 100-2000mg/L；
Polyaluminium chloride 50-150mg/L；
Newborn calcium phosphate 20-150mM.
It is above-mentioned, it should be noted that chitosan (chitosan) is also known as chitosan, is widely present by nature
Chitin (chitin) obtained by deacetylation, chemical name is Chitosan (1-4) -2- amino-B-D grapes
Sugar.
It is above-mentioned, it should be noted that be commonly called as water purification agent also known as polyaluminium chloride, abbreviation poly-aluminium, english name PAC.With alkali formula
Aluminium polychloride, spray drying aluminium polychloride belong to associated class Water purification medicament.It is a kind of polyhydroxy, the sun of body is complexed in multinuclear
Ionic inorganic polymer flocculant, solid product appearance are yellow or white solid powder, and chemical molecular formula is AL2 (OH)
NCL6-nm, in formula, 1≤n≤5, m≤10, and it is soluble easily in water, there is stronger bridge formation adsorptivity, with electrification in hydrolytic process
It learns, cohesion, the physico variation such as absorption and precipitation ultimately generates 3 (OH) 3 of AL2 (OH), to reach purification purpose.It is nontoxic, but
It is that the inside is harmful containing aluminium ion, excessive intake can lead to calcium deficiency, cause to damage to brain, accumulate in the portions such as liver,spleen,kidney
Position, interferes the digestion and absorption function of human body.
A kind of cell culture liquid decolorizer of present invention offer and discoloration method.Wherein, the cell culture liquid decolorizer is logical
It crosses using the obtained cell culture liquid decolorizer of one or more combinations in chitosan, polyaluminium chloride and newborn calcium phosphate,
It is acted on using the flocculation adsorption of chitosan, polyaluminium chloride and newborn calcium phosphate, by adding chitosan in cell culture fluid, gathering
One or more combinations in aluminium chloride and newborn calcium phosphate carry out adsorpting pigment, and then by pigment as cell removes together, from
And reach decolorizing effect, to realize in the production of the biogenetic products such as monoclonal antibody or diagnostic reagent, to the color in cell culture fluid
The abundant purifying and decoloration of element and impurity, purification efficiency is high, alleviates burden for work such as further extraction, filterings, significantly
The cost for reducing purifying and production brings huge convenience to the production, research and development and use of cell culture fluid.
In addition, in the decolorising agent, other can also be added and generates flocculate or precipitation with actual constituent phase reaction
It is sub- to can include but is not limited to aluminium polychloride, polyaluminum ferric chloride, aluminium chloride, polyacrylamide, sulfuric acid for cleanser
Iron, aluminum sulfate, bodied ferric sulfate etc..Its addition can be added according to final concentration 30-60mM, such as 30mM, 35mM,
40mM、45mM、50mM、55mM、60mM。
In addition, in the decolorising agent, EDTA can be added and play certain catharsis, EDTA also known as ethylenediamine tetrem
Acid is a kind of organic compound, chemical formula C10H16N2O8, it is white powder under normal temperature and pressure.It is it is a kind of can and Mg2+、
Ca2+、Mn2+、Fe2+The chelating agent that equal bivalent metal ions combine.Since the effect of most nucleic acid enzymes and some proteases needs
Mg2+ is wanted, therefore commonly uses the inhibitor for doing nuclease, protease；It can also be used for inhibiting effect of the heavy-metal ion removal to enzyme.On
State EDTA addition can add parts by weight be 0.5-20 part, addition can for 0.5 part, 1 part, 3 parts, 3.5 parts, 7
Part, 10 parts, 12 parts, 15 parts, 17 parts, 19 parts, 19.5 parts, 20 parts etc..
In addition, in the decolorising agent, the strong adsorbent that such as activated carbon can also be added carries out suction to fractions
Attached effect.The activated carbon of addition different-grain diameter, such as the activated carbon etc. of Nano grade can be selected according to actual needs.It adds
Entering amount can be added according to specific needs, the activated carbon to be failed again by centrifuging or filtering removal after the completion of absorption.This
Outside, the activated carbon can also be activated by the other methods such as drying or cleaning, to be reused, reduce at
This.Such as additive amount can be 50-100 parts.
In addition, in the decolorising agent, silica gel or other reagent reagents can also be added, such as can include but is not limited to
Positive silica gel, reverse phase silica gel (RP18 RP20 etc.), sephadex (LH-20), alchlor etc. play filtering or molecular sieve etc.
The material or reagent of effect.Specifically used method can be directly added into the filter material refiltered after adsorption, or for carry out
The mode of dress column filtering realizes depigmentation.
Above-mentioned, newborn calcium phosphate needs now with the current for the precipitated calcium phosphate generated by the reaction of different compounds.
Preferably, the concentration control of each component is in the cell culture fluid：
Chitosan 500-1500mg/L；
Polyaluminium chloride 80-100mg/L；
Newborn calcium phosphate 40-120mM.
Preferably,
It further include cosolvent；
The cosolvent is concentrated hydrochloric acid；
Final concentration control of the cosolvent in the cell culture fluid is 0.001-0.1mg/L.
It is above-mentioned, since chitosan powder is insoluble in water, cannot be added directly into culture solution, therefore first with ultrapure before decolorization
Chitosan stirring is configured to concentrate solution by water, with concentrated hydrochloric acid hydrotropy until completing to dissolve.
In addition, to solve the above problems, the present invention also provides a kind of cell culture fluid discoloration methods, including：
Chitosan is subjected to hydrotropy dissolving by cosolvent, obtains chitosan solution；It is prepared when carrying out culture solution decoloration
Newborn calcium phosphate；Then, take the chitosan solution, polyaluminium chloride and newborn calcium phosphate it is one or more be separately added into it is described
Culture solution to be decolourized；Wherein, the amount of the newborn calcium phosphate, chitosan, polyaluminium chloride is prepared respectively to reach default dense eventually
Degree, obtains mixed-culture medium；
The mixed-culture medium is mixed and is centrifuged to get to the cell culture fluid after decoloration.
It is above-mentioned, it, can also be before obtaining mixed-culture medium or to obtain mixed-culture medium laggard during being decolourized
The capable activated carbon that the different-grain diameter for playing certain suction-operated is added, purification on normal-phase silica gel, reverse phase silica gel, sephadex, tri-chlorination
The adsorbents such as aluminium, macroporous absorbent resin or molecular sieve, to reach certain decolorizing effect.Its addition can be existed with reality
The amount of pigment is allocated in cell culture fluid to be decolourized, for example, the upper of the 5%-10% of cell culture fluid gross mass is added
State adsorbent or molecular sieve.In addition, also column can be filled by the way that above-mentioned adsorbent or molecular sieve are carried out self filler, and then pass through preparation
Or partly prepare HPLC and carry out quick column excessively, to achieve the effect that efficiently to decolourize to cell culture fluid.
Above-mentioned, hybrid mode can be to be shaken by shaking table, or according to the amount of specific culture solution or pass through different model
Mixing plant be stirred, until uniformly until.
It is above-mentioned, centrifugation apparatus can be centrifuge, can also be needed according to actual biological product select temperature or other
Condition, such as can be ultralow temperature centrifugation or traditional vacuum.
Above-mentioned, cosolvent is hydrochloric acid, in addition it is also possible to select other shadows to product according to the product in specific culture solution
Ring smaller acid reagent, such as hydrochloric acid, phosphoric acid, acetic acid etc..
It is above-mentioned, after the cell culture fluid after obtaining the decoloration, the cell culture fluid after decoloration can also be carried out not
With the inspection of mode to realize quality control, for example, carry out visually observing experiment, ultraviolet spectrophotometry passes through OD values or turbidity
It measures its turbidity and judges clarity, or tested to cell culture fluid by high performance liquid chromatography.
Above-mentioned, the decolorising agent of addition is configured according to practical default final concentration, can also measure progress according to actual needs
Addition, until reaching clarification position, because there is also the pigment containing different content or nitrogen, ammonia etc. are miscellaneous for different cell culture fluids
Matter can carry out carrying out groping to test to the addition of decolorising agent, according to actual needs so that it is determined that the amount of being actually added into.
Preferably, described " chitosan being carried out hydrotropy dissolving by cosolvent, obtain chitosan solution " includes：
Based on default chitosan solution concentration, 0.1-5 parts of the chitosan is added to the water mixing, obtains concentrating molten
Liquid；
0.001-0.1 parts of the cosolvent is added in the concentrate solution to get to chitosan solution.
Preferably, a concentration of 3-10g/L of default chitosan solution.
Above-mentioned, the concentration of chitosan solution can be 3-10g/L, it is preferred that can be 5g/L in the present embodiment.
Preferably, described " newborn calcium phosphate is prepared when carrying out culture solution decoloration ", including：
It takes calcium chloride and sodium dihydrogen phosphate to be added to the water reaction and generates calcium phosphate, the newborn phosphorus is prepared after mixing
Sour calcium.
Preferably, described default final concentration of：
Chitosan 100-2000mg/L；
Polyaluminium chloride 50-150mg/L；
Newborn calcium phosphate 20-150mM.
Preferably, described " mixed-culture medium is mixed and is centrifuged to get to the cell culture fluid after decoloration "
Including：
The mixed solution is stirred 5-10 minutes at ambient temperature, mixing speed 50-100rpm is stirred
Mix mixed solution；
The solution that is stirred is centrifuged, centrifugal force is set as 8000-12000g, temperature 2-8, time 20-30
Minute to get to the cell culture fluid after decoloration.
It is above-mentioned, centrifugal force when centrifugation, preferably 8000g, in addition, the range of temperature may remain in 2-8 DEG C when centrifugation,
Ultralow temperature centrifuge is used to carry out centrifugal treating when necessary.To facilitate the understanding of the present invention, come with reference to embodiment further
Illustrate technical scheme of the present invention.Applicant states that the present invention illustrates that the detailed process of the present invention is set by above-described embodiment
Standby and technological process, but the invention is not limited in above-mentioned detailed process equipment and technological processes, that is, do not mean that the present invention must
Above-mentioned detailed process equipment and technological process, which must be relied on, to be implemented.Person of ordinary skill in the field is it will be clearly understood that this
Any improvement of invention, the addition of equivalence replacement and auxiliary element to each raw material of product of the present invention, the selection etc. of concrete mode,
It all falls within protection scope of the present invention and the open scope.Data and parameter in liquid processing method are cultivated in 1 embodiment 1-9 of table
1, cell culture fluid discoloration method in embodiment 1-9：
Chitosan processing and Adding Way：Based on default chitosan solution concentration, chitosan is added to the water mixing, is obtained
Concentrate solution；Cosolvent is added in the concentrate solution to get to chitosan solution；
Newborn calcium phosphate prepares and Adding Way：It takes calcium chloride and sodium dihydrogen phosphate to be added to the water reaction and generates calcium phosphate,
The newborn calcium phosphate is prepared after mixing；Also,
Step 1, it takes cell culture liquid decolorizer that preparation respectively is added in cell culture fluid to be decolourized and reaches default dense eventually
Degree, obtains mixed-culture medium；
Step 2, the mixed solution is stirred 5-10 minutes, mixing speed 50-100rpm at ambient temperature,
It obtains being stirred solution；
Step 3, the solution that is stirred being centrifuged, centrifugal force is set as 8000-12000g, 2-8 DEG C of temperature, when
Between 20-30 minutes to get to the cell culture fluid after decoloration.
The parameters such as final concentration and processing method in specific method, the different data referring in table 1 according to the embodiment are joined
Number.
2, positive controls processing method：Cell culture fluid to be decolourized is taken to be centrifuged, centrifugal force is set as 8000-
12000g, 2-8 DEG C of temperature, obtain sample to be tested at time 20-30 minute.
3, negative control group processing method：The cell culture fluid to be decolourized for taking no bleaching, not centrifuging, obtains to be measured
Sample.
4, detection method：
4.1 turbidity measure turbidity value：
Experimental facilities：WZT-2000 types photoelectric turbidimeter (the good instrument of Shanghai strength)；
Measure bottle；
Experimental method：
It takes 30mL samples to be added to measure in bottle, is put into apparatus measures slot, measures and record, concrete outcome is shown in Table 1.
4.2 vision comparison methods：
It takes sample to be tested to be respectively put into corresponding 20mL biochemistry centrifuge tube, is directly observed
Embodiment 1：
Based on the data and parameter of the present embodiment in table 1, the thin of decoloration is treated by above-mentioned cell culture fluid discoloration method
Born of the same parents' culture solution decolourizes, and the results are shown in Table 2.
Embodiment 2：
Based on the data and parameter of the present embodiment in table 1, the thin of decoloration is treated by above-mentioned cell culture fluid discoloration method
Born of the same parents' culture solution decolourizes, and the results are shown in Table 2.
Embodiment 3：
Based on the data and parameter of the present embodiment in table 1, the thin of decoloration is treated by above-mentioned cell culture fluid discoloration method
Born of the same parents' culture solution decolourizes, and the results are shown in Table 2.
Embodiment 4：
Based on the data and parameter of the present embodiment in table 1, the thin of decoloration is treated by above-mentioned cell culture fluid discoloration method
Born of the same parents' culture solution decolourizes, and the results are shown in Table 2.
Embodiment 5：
Based on the data and parameter of the present embodiment in table 1, the thin of decoloration is treated by above-mentioned cell culture fluid discoloration method
Born of the same parents' culture solution decolourizes, and the results are shown in Table 2.
Embodiment 6：
Based on the data and parameter of the present embodiment in table 1, the thin of decoloration is treated by above-mentioned cell culture fluid discoloration method
Born of the same parents' culture solution decolourizes, and the results are shown in Table 2.
Embodiment 7：
Based on the data and parameter of the present embodiment in table 1, the thin of decoloration is treated by above-mentioned cell culture fluid discoloration method
Born of the same parents' culture solution decolourizes, and the results are shown in Table 2.
Embodiment 8：
Based on the data and parameter of the present embodiment in table 1, the thin of decoloration is treated by above-mentioned cell culture fluid discoloration method
Born of the same parents' culture solution decolourizes, and the results are shown in Table 2.
Embodiment 9：
Based on the data and parameter of the present embodiment in table 1, the thin of decoloration is treated by above-mentioned cell culture fluid discoloration method
Born of the same parents' culture solution decolourizes, and the results are shown in Table 2.
Embodiment 10：
Based on the data and parameter of the present embodiment in table 1, the thin of decoloration is treated by above-mentioned cell culture fluid discoloration method
Born of the same parents' culture solution decolourizes, and the results are shown in Table 2.
Experimental result and discussion：
Testing result comparison in 2 embodiment 1-9 of table after culture solution decoloration
By above-mentioned discoloration method, embodiment 1-9 and positive controls, negative control group are respectively processed, and
It carries out turbidity value detection respectively to finally obtained sample to be tested and compares, and observation pair of the naked eyes to color and clarity
Than concrete outcome is shown in Table 2.By in table 2 it is known that culture solution handled by embodiment 1-10 is located in advance by adding decolorising agent
After reason compared with positive controls, the turbidity value of cell harvest liquid declines, and the concentration of coloring matter declines, and liquid color is by deep brown
Discoloration is light brown, and decolorizing effect is notable.
Claims (9)
1. a kind of cell culture liquid decolorizer, which is characterized in that including：
It is one or more in chitosan, polyaluminium chloride and newborn calcium phosphate；
Wherein, the concentration control of each component is in cell culture fluid：
Chitosan 100-2000mg/L；
Polyaluminium chloride 50-150mg/L；
Newborn calcium phosphate 20-150mM.
2. cell culture liquid decolorizer as described in claim 1, which is characterized in that each component is dense in the cell culture fluid
Degree controls：
Chitosan 500-1500mg/L；
Polyaluminium chloride 80-100mg/L；
Newborn calcium phosphate 40-120mM.
3. cell culture liquid decolorizer as claimed in claim 1 or 2, which is characterized in that
It further include cosolvent；
The cosolvent is concentrated hydrochloric acid；
Final concentration control of the cosolvent in the cell culture fluid is 0.001-0.1mg/L.
4. a kind of cell culture fluid discoloration method, which is characterized in that including：
Chitosan is subjected to hydrotropy dissolving by cosolvent, obtains chitosan solution；
Newborn calcium phosphate is prepared when carrying out culture solution decoloration；Then, the chitosan solution, polyaluminium chloride and newborn phosphoric acid are taken
The one or more of calcium are separately added into the culture solution to be decolourized；Wherein, prepare respectively the newborn calcium phosphate, chitosan,
The amount of polyaluminium chloride obtains mixed-culture medium to reach default final concentration；
The mixed-culture medium is mixed and is centrifuged to get to the cell culture fluid after decoloration.
5. cell culture fluid discoloration method as claimed in claim 4, which is characterized in that it is described " by chitosan by cosolvent into
Row hydrotropy dissolves, and obtains chitosan solution " include：
Based on default chitosan solution concentration, 0.1-5 parts of the chitosan is added to the water mixing, obtains concentrate solution；
0.001-0.1 parts of the cosolvent is added in the concentrate solution to get to chitosan solution.
6. cell culture fluid discoloration method as claimed in claim 5, which is characterized in that the default a concentration of 3- of chitosan solution
10g/L。
7. cell culture fluid discoloration method as claimed in claim 4, which is characterized in that described " to match when carrying out culture solution decoloration
The newborn calcium phosphate of system ", including：
It takes calcium chloride and sodium dihydrogen phosphate to be added to the water reaction and generates calcium phosphate, the newborn phosphoric acid is prepared after mixing
Calcium.
8. cell culture fluid discoloration method as claimed in claim 4, which is characterized in that described default final concentration of：
Chitosan 100-2000mg/L；
Polyaluminium chloride 50-150g/L；
Calcium phosphate 40-120mM.
9. cell culture fluid discoloration method as claimed in claim 4, which is characterized in that described " to carry out the mixed-culture medium
Mix and centrifuge to get to the cell culture fluid after decoloration " include：
The mixed solution is stirred 5-10 minutes, mixing speed 50-100rpm at ambient temperature, it is mixed to obtain stirring
Close solution；
To it is described be stirred solution and carry out centrifugation centrifugal force be set as 8000-12000g, 2-8 DEG C of temperature, time 20-30 point
Clock is to get to the cell culture fluid after the decoloration.
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Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US2305657A (en) * | 1938-03-07 | 1942-12-22 | Aehnelt Walter Robert | Decolorizing and purifying liquids |
| WO1989008718A1 (en) * | 1988-03-17 | 1989-09-21 | The Dow Chemical Company | Process for decolorizing aqueous sugar solutions via adsorbent resins, and desorption of color bodies from the adsorbent resins |
| CN102002075A (en) * | 2009-08-13 | 2011-04-06 | 楼响瑜 | Method for extracting raffinose from cottonseed wastewater |
| CN102123749A (en) * | 2008-06-23 | 2011-07-13 | 淡马锡理工学院 | A sorbent for a dialysis device |
| CN102702284A (en) * | 2012-05-23 | 2012-10-03 | 广西甙元植物制品有限公司 | Production process of high-purity sweet tea glycosides |
| CN104694010A (en) * | 2014-12-19 | 2015-06-10 | 墨江森源科技有限公司 | Preparation method of dewaxed and decolorized shellac |
| CN104884147A (en) * | 2012-10-24 | 2015-09-02 | 建新公司 | Elution of biomolecules from multi-modal resins using mes and mops as mobile phase modifiers |
| CN106319098A (en) * | 2016-09-26 | 2017-01-11 | 广西科技大学 | Sugar juice decoloring agent and sugar juice purification decoloring method |
| CN106478769A (en) * | 2016-09-23 | 2017-03-08 | 广东东阳光药业有限公司 | A kind of purification process of preliminary treatment Chinese hamster ovary celI culture fluid |
| CN107383128A (en) * | 2017-08-01 | 2017-11-24 | 江南大学 | A kind of discoloration method of steviol glycoside aqueous extract |
-
2018
- 2018-04-10 CN CN201810315766.7A patent/CN108479110A/en active Pending
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US2305657A (en) * | 1938-03-07 | 1942-12-22 | Aehnelt Walter Robert | Decolorizing and purifying liquids |
| WO1989008718A1 (en) * | 1988-03-17 | 1989-09-21 | The Dow Chemical Company | Process for decolorizing aqueous sugar solutions via adsorbent resins, and desorption of color bodies from the adsorbent resins |
| CN102123749A (en) * | 2008-06-23 | 2011-07-13 | 淡马锡理工学院 | A sorbent for a dialysis device |
| CN102002075A (en) * | 2009-08-13 | 2011-04-06 | 楼响瑜 | Method for extracting raffinose from cottonseed wastewater |
| CN102702284A (en) * | 2012-05-23 | 2012-10-03 | 广西甙元植物制品有限公司 | Production process of high-purity sweet tea glycosides |
| CN104884147A (en) * | 2012-10-24 | 2015-09-02 | 建新公司 | Elution of biomolecules from multi-modal resins using mes and mops as mobile phase modifiers |
| CN104694010A (en) * | 2014-12-19 | 2015-06-10 | 墨江森源科技有限公司 | Preparation method of dewaxed and decolorized shellac |
| CN106478769A (en) * | 2016-09-23 | 2017-03-08 | 广东东阳光药业有限公司 | A kind of purification process of preliminary treatment Chinese hamster ovary celI culture fluid |
| CN106319098A (en) * | 2016-09-26 | 2017-01-11 | 广西科技大学 | Sugar juice decoloring agent and sugar juice purification decoloring method |
| CN107383128A (en) * | 2017-08-01 | 2017-11-24 | 江南大学 | A kind of discoloration method of steviol glycoside aqueous extract |
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